Fig 2: MLH1 loss caused by DNMT3A/B upregulation is mediated through the PP2A-Rb-E2F pathway.a Immunoprecipitation performed with Capturem Protein A columns. Organoid lysates from WT mice were incubated with 1 µg of PPP2R1A antibody for 10 min. The antibody–lysate complex was applied to equilibrate Protein A spin columns. The eluted fraction was then subjected to SDS-PAGE to confirm the presence of PP2A B subunit antigen (52 kDa, detected by Cell Signaling #2290). Images are representative of one biological independent sample for each group. b Volcano plot for the comparison of RNA-seq data of Lgr5-EGFP-CreERT2; Ppp2r1aflox/flox intestinal organoids treated with and without DMBA and tamoxifen for 50 days. Differently expressed genes (fold change >2 and FDR < 0.05) are denoted in red or green. Plot is representative of three biological independent samples for each group. c Intersection of ppp2r1a interaction protein in a and differently expressed genes in b data (IP protein data are available in Supplementary Data 1 and RNA-seq data are available in GSE120241 with NIH/NCBI at GEO dataset). d The top 3 significant gene ontology categories from the 58 interacted proteins identified by GSEA (whole gene ontology category is listed in Supplementary Table 4). e Venn diagram showing overlap (gene numbers) among genes associated with regulation of TP53 activity, cell cycle, and RNA polymerase II transcription. The intersection contained four genes, including hdac2, ppp2cb, akt1, and rb1. Western blot analysis of f CT26 transfected with the indicated shRNAs, and treated with g vehicle control or 5-azacytidine (AZA, 1 μM for 1 days). Blots are representative of three biological independent samples for each group. h Human tissue array containing MSI and MSS colorectal tumours was assayed for p-Rb (n = 141, P = 0.0132) and p-HDAC2 (n = 138, P = 0.02) levels by immunohistochemistry. (Left) Representative pictures showing increased p-Rb and p-HDAC2 levels in MSI compared to MSS. Arrows indicate positive signals. Bar = 25 μm. (Right) Quantitative data are shown. P value was determined by two-sided Mann–Whitney U-test. Source data are provided as a Source data file.

Fig 3: Western blot analysis.Western blot analysis of JMJD3 expression and the downstream signaling pathway in the SAS and Cal 27 cell lines. The protein expression profiles of JMJD3, p21, Rb and pRb were examined in the presence or absence of GSK-J4 treatment in the oral tongue squamous cell carcinoma cells by Western blotting.

Fig 4: pRB and CYCA2 positive cells (arrows) can be seen in the eye section of an 8-week human embryo-8-w.HE (a), retinoblastoma-Rb (b), normal human eye-NHE (c) and epithelioid melanoma-EM (d); r—retina, *—choroidea, tm—tumor tissue. Co-localization of pRB and CYCA2 positive cells (arrowhead in the inset of magnification frame). Double immunofluorescence staining to pRB (first column), CYCA2 (second column), DAPI (third column), merge (fourth column). Magnification frames provide detail with positive cells (arrows) or co-localization (arrowheads). Scale bar 25 μm.

Fig 5: HDAC4 knockdown induces G1/S arrest in EC cells(A and C) Flow cytometry analysis of indicated EC cells transfected with HDAC4-specific siRNAs or a control siRNA; G1-phase cells are significantly increased while S-phase cells are reduced in HDAC4 knockdown (B) KYSE30 and (D) EC/CUHK1 cells, * P <0.05; (E) HDAC4 down-regulation increases the expression of p21 and p27 proteins while reduces pRb, CDK2 and CDK4 in EC cells.